PUL ID

PUL0041

PubMed

9023916, Appl Environ Microbiol. 1997 Feb;63(2):355-63. doi: 10.1128/AEM.63.2.355-363.1997.

Characterization method

Southern Blot, enzyme activity assay

Genomic accession number

U61727

Nucelotide position range

1-3733

Substrate

cellobiose

Loci

casRAB

Species

Klebsiella oxytoca/571

Degradation or Biosynthesis

degradation

Gene Name

Locus Tag

Protein ID

Gene Position

GenBank Contig Range

EC Number

casR - AAB51562.1 0 - 291 (+) U61727.1:1-292 -
casA - AAB51563.1 455 - 2321 (+) U61727.1:456-2322 -
casB - AAB51564.1 2338 - 3733 (+) U61727.1:2339-3734 -

Cluster number

1

Gene name

Gene position

Gene type

Found by CGCFinder?

- 1 - 291 (+) CDS No
- 456 - 2321 (+) TC: gnl|TC-DB|P08722|4.A.1.2.2 Yes
- 2339 - 3733 (+) CAZyme: GH1 Yes

PUL ID

PUL0041

PubMed

9023916, Appl Environ Microbiol. 1997 Feb;63(2):355-63. doi: 10.1128/AEM.63.2.355-363.1997.

Title

Cloning of cellobiose phosphoenolpyruvate-dependent phosphotransferase genes: functional expression in recombinant Escherichia coli and identification of a putative binding region for disaccharides.

Author

Lai X, Davis FC, Hespell RB, Ingram LO

Abstract

Genomic libraries from nine cellobiose-metabolizing bacteria were screened for cellobiose utilization. Positive clones were recovered from six libraries, all of which encode phosphoenolpyruvate:carbohydrate phosphotransferase system (PTS) proteins. Clones from Bacillus subtilis, Butyrivibrio fibrisolvens, and Klebsiella oxytoca allowed the growth of recombinant Escherichia coli in cellobiose-M9 minimal medium. The K. oxytoca clone, pLOI1906, exhibited an unusually broad substrate range (cellobiose, arbutin, salicin, and methylumbelliferyl derivatives of glucose, cellobiose, mannose, and xylose) and was sequenced. The insert in this plasmid encoded the carboxy-terminal region of a putative regulatory protein, cellobiose permease (single polypeptide), and phospho-beta-glucosidase, which appear to form an operon (casRAB). Subclones allowed both casA and casB to be expressed independently, as evidenced by in vitro complementation. An analysis of the translated sequences from the EIIC domains of cellobiose, aryl-beta-glucoside, and other disaccharide permeases allowed the identification of a 50-amino-acid conserved region. A disaccharide consensus sequence is proposed for the most conserved segment (13 amino acids), which may represent part of the EIIC active site for binding and phosphorylation.